{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T20:35:44Z","timestamp":1761510944764},"reference-count":34,"publisher":"American Society for Microbiology","issue":"11","license":[{"start":{"date-parts":[[2010,11,1]],"date-time":"2010-11-01T00:00:00Z","timestamp":1288569600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2010,11]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>Eleven different lyssavirus species, four of which occur on the African continent, are presently recognized. These viruses cause rabies, the burden of which is highest in the developing world, where routine laboratory diagnosis is often not available. From an epidemiological and control perspective, it is necessary that diagnostic methods detect the diversity of lyssaviruses present in different regions of the world. A published and widely used heminested reverse transcription-PCR (hnRT-PCR) was evaluated for its ability to detect a panel of diverse African lyssaviruses. Due to the limitations experienced for this assay, an alternative hnRT-PCR was developed. The new assay was found to be accurate and sensitive in the detection of African lyssavirus RNA in a variety of clinical specimens. The assay was further adapted to a real-time PCR platform to allow rapid, one-step, quantitative, and single-probe detection, and an internal control for the verification of sample preparation was included. The limit of detection of the real-time PCR assay was 10 RNA copies per reaction, with inter- and intra-assay variability below 4%. Subsequently, in demonstrating utility, both assays were successfully applied to antemortem rabies diagnosis in humans. We believe that the quantitative real-time PCR assay could find application as a routine confirmatory test for rabies diagnosis in the future and that it will serve as a valuable research tool in the biology of African lyssaviruses. Alternatively, the hnRT-PCR assay can be used in laboratories that do not have access to expensive real-time PCR equipment for sensitive diagnosis of lyssaviruses.<\/jats:p>","DOI":"10.1128\/jcm.01256-10","type":"journal-article","created":{"date-parts":[[2010,9,2]],"date-time":"2010-09-02T02:49:35Z","timestamp":1283395775000},"page":"3949-3955","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":52,"title":["Improved PCR Methods for Detection of African Rabies and Rabies-Related Lyssaviruses"],"prefix":"10.1128","volume":"48","author":[{"given":"Jessica","family":"Coertse","sequence":"first","affiliation":[{"name":"Department of Microbiology and Plant Pathology, Faculty of Natural and Agricultural Sciences, University of Pretoria, Pretoria, 0001, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacqueline","family":"Weyer","sequence":"additional","affiliation":[{"name":"National Institute for Communicable Diseases of the National Health Laboratory Services, Sandringham, 2131, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Louis H.","family":"Nel","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Plant Pathology, Faculty of Natural and Agricultural Sciences, University of Pretoria, Pretoria, 0001, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wanda","family":"Markotter","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Plant Pathology, Faculty of Natural and Agricultural Sciences, University of Pretoria, Pretoria, 0001, South Africa"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"445","year":"1996","unstructured":"Aubert, M. F. A. 1996. Methods for the calculation of titers, p. 445-447. In F. X. Meslin, M. M. Kaplan, and H. Koprowski (ed.), Laboratory techniques in rabies, 4th ed. World Health Organization, Geneva, Switzerland.","journal-title":"Laboratory techniques in rabies"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-0934(02)00062-9"},{"key":"e_1_3_2_4_2","first-page":"35","volume":"49","year":"2007","unstructured":"Blumberg, L., J. Weyer, J. Frean, and G. A. Ogunbanjo. 2007. Rabies: an evidence-based approach to management. S. Afr. Fam. Pract. 49 : 35-41.","journal-title":"S. Afr. Fam. Pract."},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0912.030374"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2008.112797"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1086\/592969"},{"key":"e_1_3_2_8_2","first-page":"88","year":"1996","unstructured":"Dean, D. J., M. K. Abelseth, and P. Atansiu. 1996. The fluorescent antibody test, p. 88-95. In F. X. Meslin, M. M Kaplan, and H. Koprowski (ed.), Laboratory techniques in rabies, 4th ed. World Health Organization, Geneva, Switzerland.","journal-title":"Laboratory techniques in rabies"},{"key":"e_1_3_2_9_2","first-page":"95","volume":"41","year":"1999","unstructured":"Hall, T. A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95\/98\/NT. Nucleic Acids Symp. Ser. 41 : 95-98.","journal-title":"Nucleic Acids Symp. Ser."},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.35.11.2762-2766.1997"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00705-005-0785-0"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.42.1.299-306.2004"},{"key":"e_1_3_2_13_2","unstructured":"International Committee on Taxonomy of Viruses. 2009. ICTV official taxonomy: updates since the 8th report. Vertebrate. International Committee on Taxonomy of Viruses. http:\/\/talk.ictvonline.org\/files\/ictv_official_taxonomy_updates_since_the_8th_report\/m\/vertebrate_2008\/1208.aspx\n."},{"key":"e_1_3_2_14_2","first-page":"80","year":"1996","unstructured":"Koprowski, H. 1996. The mouse inoculation test, p. 80-85. In F. X. Meslin, M. M. Kaplan, and H. Koprowski (ed.), Laboratory techniques in rabies, 4th ed. World Health Organization, Geneva, Switzerland.","journal-title":"Laboratory techniques in rabies"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2010.01.018"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1412.080750"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2005.03.008"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-1702(03)00217-X"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1301.060810"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.virusres.2008.02.001"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid1212.060514"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.21547"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/0166-0934(95)01907-3"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2199-7-5"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1080\/10408410701647602"},{"key":"e_1_3_2_26_2","first-page":"161","year":"2007","unstructured":"Nel, L. H., and C. E. Rupprecht. 2007. Emergence of Lyssaviruses in the Old World: the case of Africa, p. 161-193. In J. E. Childs, J. S. Mackenzie, and J. A. Richt (ed.), Wildlife and emerging zoonotic diseases: the biology, circumstances and consequences of cross-species transmission. Springer-Verlag, Berlin, Germany.","journal-title":"Wildlife and emerging zoonotic diseases: the biology, circumstances and consequences of cross-species transmission"},{"key":"e_1_3_2_27_2","first-page":"297","volume":"65","year":"1998","unstructured":"Nel, L. H., J. Bingham, J. A. Jacobs, and J. B. Jaftha. 1998. A nucleotide-specific polymerase chain reaction assay to differentiate rabies virus biotypes in South Africa. Onderstepoort J. Vet. Res. 65 : 297-303.","journal-title":"Onderstepoort J. Vet. Res."},{"key":"e_1_3_2_28_2","first-page":"87","year":"2004","unstructured":"Pfaffl, M. W. 2004. Quantification strategies in real-time PCR, p. 87-120. In S. A. Bustin (ed.), A-Z of quantitative PCR. La Jolla International University Line, La Jolla, CA.","journal-title":"A-Z of quantitative PCR"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2003.11.015"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1586\/erm.09.85"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jviromet.2008.05.004"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.43.6.2786-2792.2005"},{"key":"e_1_3_2_33_2","first-page":"86","volume":"16","year":"2007","unstructured":"Weyer, J., and L. Blumberg. 2007. Rabies: challenge of diagnosis in resource-poor countries. Infect. Dis. J. Pak. 16 : 86-88.","journal-title":"Infect. Dis. J. Pak."},{"key":"e_1_3_2_34_2","unstructured":"Expert consultation on rabies technical report series 931. 2005"},{"key":"e_1_3_2_35_2","unstructured":"World Organisation for Animal Health. 2006. Manual of diagnostic tests and vaccines for terrestrial animals 5th ed. World Organisation for Animal Health Paris France. http:\/\/www.oie.int\/eng\/normes\/mmanual\/a_00044.htm\n."}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.01256-10","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.01256-10","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,5]],"date-time":"2022-03-05T08:37:16Z","timestamp":1646469436000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JCM.01256-10"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2010,11]]},"references-count":34,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2010,11]]}},"alternative-id":["10.1128\/JCM.01256-10"],"URL":"https:\/\/doi.org\/10.1128\/jcm.01256-10","relation":{},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2010,11]]},"assertion":[{"value":"2010-06-21","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2010-08-22","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2010-11-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}